Speech Technologies for African Languages: Example of a Multilingual Calculator for Education

International audienceThis paper presents our achievements after 18 months of the ALFFA project dealing with African languages technologies. We focus on a multilingual calculator (Android app) that will be demonstrated during the Show and Tell session

Site Specific Knock-In Genome Editing in Human HSCs Using Baboon Envelope gp Pseudotypedviral Derived “Nanoblades” Loaded with Cas9/sgRNA Combined with Donor Encoding AAV-6

Programmable nucleases have enabled rapid and accessible genome engineering in eukaryotic cells and living organisms. Here, we have designed ?Nanoblades?, a new technology that will deliver a genomic cleaving agent into cells. These are genetically modified Murine Leukemia Virus (MLV) or HIV derived virus like particle (VLP), in which the viral structural protein Gag has been fused to the Cas9. These VLPs are thus loaded with Cas9 protein together with the guide RNAs. Thus, nanoblades are devoid of any viral-derived genetic material. Highly efficient gene editing was obtained in cell lines, IPS cells and primary mouse and human cells (Mangeot et al. Nature Communication, 2019). However, their delivery into target cells can be technically challenging when working with primary immune cells. Now we showed that nanoblades were remarkably efficient for entry into human T, B and hematopoietic stem cells thanks to their surface co-pseudotyping with baboon retroviral and VSVG envelope glycoproteins. We were able to induce efficient, transient and very rapidlygenome-editing in human induced pluripotent stem cells reaching up to 70% in the empty spiracles homeobox 1 (EMX1) and muscular dystrophy (MD) gene locus. A brief nanoblade incubation of primary human T and B cells resulted in 40% and 20% editing of the Wiskott-Aldrich syndrome (WAS) gene locus, while hematopoietic stem cells treated for 18 h with nanoblades allowed 30-40% gene editing in the WAS gene locus and up to 80% for the Myd88 genomic target. Moreover, for the HIV- and MLV-derived nanoblades no cell toxicity and low to undetectable off-target effects were demonstrated.Finally, we also treated hHSCs with nanoblades in combination with an AAV-6 donor encoding vector resulting in over 20% of stable expression cassette knock-in into the WAS gene locus. Currently, we are evaluating these gene modified HSCs for their long-term reconstitution of NOD/SCIDgC-/- mice.Summarizing, this new technology is simple to implement in any laboratory, shows high flexibility for different targets including primary immune cells of murine and human origin, is relatively inexpensive and therefore have important prospects for basic and clinical translation in the area of gene therapy.Fil: Gutierrez, Alejandra. Inserm; FranciaFil: Abrey Recalde, Maria Jimena. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Hospital Italiano. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional E Ingenieria Biomedica.; Argentina. Inserm; FranciaFil: Mangeot, Philippe E.. Inserm; FranciaFil: Costa, Caroline. Inserm; FranciaFil: Bernandin, Ornellie. Inserm; FranciaFil: Fusil, Floriane. Inserm; FranciaFil: Froment, Gisèle. Inserm; FranciaFil: Martin, Francisco. Inserm; FranciaFil: Bellabdelah, Karim. Universidad de Granada; EspañaFil: Ricci, Emiliano P.. Inserm; FranciaFil: Ayuso, Eduard. Universite de Nantes; FranciaFil: Cosset, François loic. Inserm; FranciaFil: Verhoeyen, Els. Inserm; FranciaAmerican Society of Cell and Gene Therapy 22nd Annual MettingWashingtonEstados UnidosAmerican Society of Cell and Gene Therap

Terminal NK cell maturation is controlled by concerted actions of T-bet and Zeb2 and is essential for melanoma rejection

Natural killer (NK) cell maturation is a tightly controlled process that endows NK cells with functional competence and the capacity to recognize target cells. Here, we found that the transcription factor (TF) Zeb2 was the most highly induced TF during NK cell maturation. Zeb2 is known to control epithelial to mesenchymal transition, but its role in immune cells is mostly undefined. Targeted deletion of Zeb2 resulted in impaired NK cell maturation, survival, and exit from the bone marrow. NK cell function was preserved, but mice lacking Zeb2 in NK cells were more susceptible to B16 melanoma lung metastases. Reciprocally, ectopic expression of Zeb2 resulted in a higher frequency of mature NK cells in all organs. Moreover, the immature phenotype of Zeb2(-/-) NK cells closely resembled that of Tbx21(-/-) NK cells. This was caused by both a dependence of Zeb2 expression on T-bet and a probable cooperation of these factors in gene regulation. Transgenic expression of Zeb2 in Tbx21(-/-) NK cells partially restored a normal maturation, establishing that timely induction of Zeb2 by T-bet is an essential event during NK cell differentiation. Finally, this novel transcriptional cascade could also operate in human as T-bet and Zeb2 are similarly regulated in mouse and human NK cells

Nanoblades allow high-level genome editing in murine and human organoids

Genome engineering has become more accessible thanks to the CRISPR-Cas9 gene-editing system. However, using this technology in synthetic organs called “organoids” is still very inefficient. This is due to the delivery methods for the CRISPR-Cas9 machinery, which include electroporation of CRISPR-Cas9 DNA, mRNA, or ribonucleoproteins containing the Cas9-gRNA complex. However, these procedures are quite toxic for the organoids. Here, we describe the use of the “nanoblade (NB)” technology, which outperformed by far gene-editing levels achieved to date for murine- and human tissue-derived organoids. We reached up to 75% of reporter gene knockout in organoids after treatment with NBs. Indeed, high-level NB-mediated knockout for the androgen receptor encoding gene and the cystic fibrosis transmembrane conductance regulator gene was achieved with single gRNA or dual gRNA containing NBs in murine prostate and colon organoids. Likewise, NBs achieved 20%–50% gene editing in human organoids. Most importantly, in contrast to other gene-editing methods, this was obtained without toxicity for the organoids. Only 4 weeks are required to obtain stable gene knockout in organoids and NBs simplify and allow rapid genome editing in organoids with little to no side effects including unwanted insertion/deletions in off-target sites thanks to transient Cas9/RNP expression

IRGM Is a Common Target of RNA Viruses that Subvert the Autophagy Network

Autophagy is a conserved degradative pathway used as a host defense mechanism against intracellular pathogens. However, several viruses can evade or subvert autophagy to insure their own replication. Nevertheless, the molecular details of viral interaction with autophagy remain largely unknown. We have determined the ability of 83 proteins of several families of RNA viruses (Paramyxoviridae, Flaviviridae, Orthomyxoviridae, Retroviridae and Togaviridae), to interact with 44 human autophagy-associated proteins using yeast two-hybrid and bioinformatic analysis. We found that the autophagy network is highly targeted by RNA viruses. Although central to autophagy, targeted proteins have also a high number of connections with proteins of other cellular functions. Interestingly, immunity-associated GTPase family M (IRGM), the most targeted protein, was found to interact with the autophagy-associated proteins ATG5, ATG10, MAP1CL3C and SH3GLB1. Strikingly, reduction of IRGM expression using small interfering RNA impairs both Measles virus (MeV), Hepatitis C virus (HCV) and human immunodeficiency virus-1 (HIV-1)-induced autophagy and viral particle production. Moreover we found that the expression of IRGM-interacting MeV-C, HCV-NS3 or HIV-NEF proteins per se is sufficient to induce autophagy, through an IRGM dependent pathway. Our work reveals an unexpected role of IRGM in virus-induced autophagy and suggests that several different families of RNA viruses may use common strategies to manipulate autophagy to improve viral infectivity

« 20 janvier 1800, À qui parles-tu ? » Joseph Joubert et l'écriture des carnets

Joubert's notebooks are far from being a book in any normal sense of the world. The several editions which have apeared since the last century have been selections of Joubert's writings, which imposed an order in the writer' s pages. By examining what Joubert was experimenting with in this original method of not writing a book, we can see that he was in fact inventing a new kind of space for writing, a space which is precisely that of the notebook. We can see too what kind of potential reader he was writing for.Mangeot Philippe. « 20 janvier 1800, À qui parles-tu ? » Joseph Joubert et l'écriture des carnets. In: Littérature, n°80, 1990. Carnets, cahiers. pp. 71-85

Etude expérimentale et développement numérique d'une modélisation des phénomènes physicochimiques dans un propulseur hybride spatial

La propulsion hybride utilise classiquement un comburant liquide (ou gazeux) injecté dans une chambre de combustion qui contient le carburant à l'état solide. La flamme de diffusion, qui apparait à la rencontre des deux flux de matière, est autoentretenue par la pyrolyse du carburant consécutive à l apport de chaleur produite par la combustion. Afin d'améliorer les performances de ce type de propulsion, il est nécessaire de bien comprendre le couplage physicochimique des phénomènes. Le couple d'ergols polyéthylène/mélange gazeux dioxygène et diazote a été choisi pour cette étude. Les caractéristiques du polyéthylène ont été déterminées par des analyses physicochimiques, elles permettent de mettre en évidence un effet de la pression et de la nature de l'atmosphère sur la composition des produits de pyrolyse. Un banc d'essais de combustion avec une instrumentation a permis de caractériser le comportement du polyéthylène en situation réelle. Les données acquises ont été analysées afin d'obtenir des grandeurs physiques pertinentes à comparer avec des résultats de simulations. Pour effectuer des simulations de chambre de combustion de propulseur hybride, le développement d'un modèle numérique instationnaire et bidimensionnel a débuté. De nombreux cas test "académiques" sont présentés et ont confirmés la bonne implémentation des méthodes numériques de résolution et des équations physiques et chimiques. Cependant, lors des simulations de la chambre de combustion complète, une divergence de pression est apparue dont les causes ont été activement recherchées.The hybrid space propulsion classically employs a liquid (or gaseous) oxidizer injected into the combustion chamber which contains the solid reducer. The diffusion flame, which appears at the confluence of the oxidizer and reducer mater fluxes, is auto-entertained by the fuel pyrolysis piloted by the heat which is provided to it by the combustion. In order to increase performances of this propulsion system, it is necessary to well understand the coupling effect of the physical and chemical phenomena. The propellants couple polyethylene/gaseous oxygen and nitrogen mixture has been chosen for this study. Properties of the polyethylene have been determined by several chemical analyses, showing that there are a pressure and atmosphere nature effects on the pyrolysis chemical composition. A test bench with instrumentation allowed to characterize the behavior of the polyethylene in real situation. Data acquired have been analyzed in order to obtain physical variables relevant for comparing the numerical simulations results. To undertake simulation of the combustion chamber of hybrid rocket, a new numerical model has been developed. Numerous "academic" test cases are presented and confirmed a good implementation of the numerical method and of the physical and chemical models. Nevertheless, during simulations of hybrid combustion chamber, a pressure divergence has appeared thus causes have been actively investigated.ORLEANS-SCD-Bib. electronique (452349901) / SudocSudocFranceF

Micro and Full-Scale Experiments on Hybrid Rocket Fuel and Prelude to Hybrid Rocket Combustor Simulations

International audienc